Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis belt
Improved methods for the detection and characterization of carried Neisseria meningitidis isolates are needed. We evaluated a multiplex PCR algorithm for the detection of a variety of carriage strains in the meningitis belt. To further improve the sensitivity and specificity of the existing PCR assa...
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
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| Format: | Journal article |
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Public Library of Science
2018
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| _version_ | 1826277376857735168 |
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| author | Diallo, K Coulibaly, MD Rebbetts, LS Harrison, OB Lucidarme, J Gamougam, K Tekletsion, YK Bugri, A Toure, A Issaka, B Dieng, M Trotter, C Collard, JM Sow, SO Wang, X Mayer, LW Borrow, R Greenwood, BM Maiden, MCJ Manigart, O |
| author_facet | Diallo, K Coulibaly, MD Rebbetts, LS Harrison, OB Lucidarme, J Gamougam, K Tekletsion, YK Bugri, A Toure, A Issaka, B Dieng, M Trotter, C Collard, JM Sow, SO Wang, X Mayer, LW Borrow, R Greenwood, BM Maiden, MCJ Manigart, O |
| author_sort | Diallo, K |
| collection | OXFORD |
| description | Improved methods for the detection and characterization of carried Neisseria meningitidis isolates are needed. We evaluated a multiplex PCR algorithm for the detection of a variety of carriage strains in the meningitis belt. To further improve the sensitivity and specificity of the existing PCR assays, primers for gel-based PCR assays (sodC, H, Z) and primers/probe for real-time quantitative PCR (qPCR) assays (porA, cnl, sodC, H, E, Z) were modified or created using Primer Express software. Optimized multiplex PCR assays were tested on 247 well-characterised carriage isolates from six countries of the African meningitis belt. The PCR algorithm developed enabled the detection of N. meningitidis species using gel-based and real-time multiplex PCR targeting porA, sodC, cnl and characterization of capsule genes through sequential multiplex PCR assays for genogroups (A, W, X, then B, C, Y and finally H, E and Z). Targeting both porA and sodC genes together allowed the detection of meningococci with a sensitivity of 96% and 89% and a specificity of 78% and 67%, for qPCR and gel-based PCR respectively. The sensitivity and specificity ranges for capsular genogrouping of N. meningitidis are 67% - 100% and 98%-100% respectively for gel-based PCR and 90%-100% and 99%-100% for qPCR. We developed a PCR algorithm that allows simple, rapid and systematic detection and characterisation of most major and minor N. meningitidis capsular groups, including uncommon capsular groups (H, E, Z). |
| first_indexed | 2024-03-06T23:27:59Z |
| format | Journal article |
| id | oxford-uuid:6b0bf58a-5c88-42bd-b225-f1d627c7cb40 |
| institution | University of Oxford |
| last_indexed | 2024-03-06T23:27:59Z |
| publishDate | 2018 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | oxford-uuid:6b0bf58a-5c88-42bd-b225-f1d627c7cb402022-03-26T19:01:17ZDevelopment of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis beltJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:6b0bf58a-5c88-42bd-b225-f1d627c7cb40Symplectic Elements at OxfordPublic Library of Science2018Diallo, KCoulibaly, MDRebbetts, LSHarrison, OBLucidarme, JGamougam, KTekletsion, YKBugri, AToure, AIssaka, BDieng, MTrotter, CCollard, JMSow, SOWang, XMayer, LWBorrow, RGreenwood, BMMaiden, MCJManigart, OImproved methods for the detection and characterization of carried Neisseria meningitidis isolates are needed. We evaluated a multiplex PCR algorithm for the detection of a variety of carriage strains in the meningitis belt. To further improve the sensitivity and specificity of the existing PCR assays, primers for gel-based PCR assays (sodC, H, Z) and primers/probe for real-time quantitative PCR (qPCR) assays (porA, cnl, sodC, H, E, Z) were modified or created using Primer Express software. Optimized multiplex PCR assays were tested on 247 well-characterised carriage isolates from six countries of the African meningitis belt. The PCR algorithm developed enabled the detection of N. meningitidis species using gel-based and real-time multiplex PCR targeting porA, sodC, cnl and characterization of capsule genes through sequential multiplex PCR assays for genogroups (A, W, X, then B, C, Y and finally H, E and Z). Targeting both porA and sodC genes together allowed the detection of meningococci with a sensitivity of 96% and 89% and a specificity of 78% and 67%, for qPCR and gel-based PCR respectively. The sensitivity and specificity ranges for capsular genogrouping of N. meningitidis are 67% - 100% and 98%-100% respectively for gel-based PCR and 90%-100% and 99%-100% for qPCR. We developed a PCR algorithm that allows simple, rapid and systematic detection and characterisation of most major and minor N. meningitidis capsular groups, including uncommon capsular groups (H, E, Z). |
| spellingShingle | Diallo, K Coulibaly, MD Rebbetts, LS Harrison, OB Lucidarme, J Gamougam, K Tekletsion, YK Bugri, A Toure, A Issaka, B Dieng, M Trotter, C Collard, JM Sow, SO Wang, X Mayer, LW Borrow, R Greenwood, BM Maiden, MCJ Manigart, O Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis belt |
| title | Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis belt |
| title_full | Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis belt |
| title_fullStr | Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis belt |
| title_full_unstemmed | Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis belt |
| title_short | Development of a PCR algorithm to detect and characterize Neisseria meningitidis carriage isolates in the African meningitis belt |
| title_sort | development of a pcr algorithm to detect and characterize neisseria meningitidis carriage isolates in the african meningitis belt |
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